For informations about installation of the image analysis tool, access the “Simulator” tab

 

Step 1: Import the image of Lignin stained tissue section into image analysis software.

 

 

The image will open in a new pop up window as shown.

 

Step 2: To split the section into different channels follow, image ---> color ---> split channels.

 

 

A new pop window will open, which will show the tissue section in three channels: green, blue and red.

 

 

Step 3: Select any one of the image that will be easier to analyze (Select a clear image). Here tissue section in green is selected for further analysis.

 

 

Step 4: Then adjust threshold to find the intensity of lignin staining. Follow,  Image --->  Adjust --->  Threshold

 

 

While adjusting the threshold, select the lignin stained region of the imported image.

 

 

Step 5: Using the free hand selection tool, select the stained region of the imported image.

 

 

Step 6: Set measurements to find the intensity of lignin stained region. Follow Analyze ---> Set Measurements. Select the options Area, Integrated density, Area Fraction, Limited to threshold and Display Label in the Menu. Then click OK to analyze the image.

 

 

Step 7:  To quantify the lignin stained area in the imported image, follow Analyze ---> Measure

 

 

A new pop up window will show the results of the tissue section.

 

Result Interpretation:

 

Percentage Area indicates the amount of lignin in the selected stained region. Here, 23 percent of the total selected region (Total Area 87181) is lignin.

 

 

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